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OBJECTIVE To demonstrate the long-or short-term impacts of neonatal Pxr(pregnane X receptor) agonists exposureon DMEs (drug metabolism enzymes) expression in adulthood. METHODS C57BL/6 mice(day 5,postnatal)were injected with different doses(0,50,100,150,200 mg·kg-1·d-1, constitutive 4 d)of PCN(pregnenolone-16a-carbonitrile).Mice at different ages(day 5,10,15,25,postna-tal)were administrated with 200 mg·kg-1·d-1PCN in constitutive 4 d.All mice were sacrificed at day 60 after birth. Liver samples were collected for detecting the expression of Pxr target genes. RESULTS Compared with vehicle group, the significant inductions of Cyp2b10, Cyp3a11 and Pxrwere observed in high dose groups (150, 200 mg·kg-1·d-1, 5-8 d after birth) both in male and female mice (n=4-9/group,P<0.05).Furthermore,high dose groups(200 mg·kg-1·d-1,5-8 d after birth)were found to have higher mRNA expression levels of Cyp2a4,Ugt1a1,Abcc4,and Oatpla4 in female mice,while Papss2 in male mice compared with vehicle groups (n= 4-9/group, P<0.05). Interestingly, a decreased mRNA expression of Sult2a1 was identified in 200 (5-8 d) groups (n=4-9/group, P<0.05). Consistent with these results, the protein expression of Cyp3a11 was only increased in 200 (5-8 d) groups compared with the vehicle groups(n=3/group,P<0.05).Importantly,the persistent impacts on DMEs only occurred in day 5 and day 25 treatment groups,not day 10 and day 15 groups(n=4/group).CONCLUSION Neonatal Pxr activation has a long-term effect on the expression of DMEs in C57BL/6 mice.Dose and treatment exposure time are two key factors involved in this permanent alteration procedure.
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OBJECTIVE To explore the effects of perinatal inflammation on the expression of proin-flammatory cytokines and DMEs (drug metabolism enzymes) in offspring mice. METHODS C57BL/6 maternal mice were administrated with single dose 100 μg·kg-1LPS(lipopolysaccharide)or saline(vehicle) during gestation (day 10 after fertilization). Offspring mice were sacrificed at 30 d after birth and liver samples were collected.Real-time PCR was adopted to test the mRNA expression of proinflammatory cytokines (Nrlp3 and IL-1β), nuclear receptors (Pxr and Car), and DMEs (Cyp3a11, 2b10, 1a2, and Ugt1a1).RESULTS Gender different expression of candidate genes was observed.The expression of Car,in the maternal injection of LPS groups,was significantly decreased in both female and male offspring (n=3-8/group, P<0.01). Concomitantly, a significantly lower expression of Cyp3a11 was found in both female and male offspring (P<0.01, P<0.05, respectively). Furthermore, the expression of Ugt1a1 was reduced in male offspring following maternal administration of LPS (P<0.01). In male offspring, Nrlp3 expression was specially decreased(P<0.05).Interestingly,there was an approximately 66% reduction in mRNA level of Cyp1a2 in female offspring (P<0.01), while in male offspring Cyp1a2 expression showed an increased trend (P>0.05) compared with vehicle group. The expression of Pxr, Cyp2b10, and IL-1β was no difference between LPS treatment group and vehicle group(P>0.05).CONCLUSION Maternal LPS administration affects the expression of proinflammatory cytokines, nuclear receptors and DMEs in mouse offspring.
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Objective: To observe the action characteristics of ferulic acid and isoferulic acid on cytochrome P450 isoenzymes1A1 and 3A4 (CYP1A1, CYP3A4) in HepG2 cells. Methods: MTT colorimetry was used to investigate the effects of ferulic acid and isoferulic acid at different concentration on the proliferation of in vitro cultured HepG2 cells. Flow cytometry was used to determine the effects of ferulic acid and isoferulic acid on cell cycle of human liver HepG2 cells; The expression of CYP1A1 and CYP3A4 mRNA was tested by real time quantitative PCR technology after drug treatment and then CYP3A4 protein expression was detected by protein immunoblot method (Western blotting). Results: HepG2 cells were inhibited by ferulic acid and isoferulic acid after 48 h action, with a clear dose-response relationship; Two drugs (50 μg/mL) blocked HepG2 cell cycle in G2-M phase; All of the concentration (100, 50, and 25 μg/mL) of the two drugs were CYP1A1 and CYP3A4 mRNA inhibitors; After cells were disposed by the two kinds of drugs (50 μg/mL) with 48 h, protein expression was significantly lower than that in the control group; And compared with the control group, the expression amounts of ferulic acid and isoferulic acid were 0.57 and 0.39. Conclusion: The proliferation of HepG2 cells is inhibited by the two drugs, and the cell cycle was arrested at G2/M phase, which that may be one of its mechanisms, so as to inhibit the mRNA expression of CYP1A1 and CYP3A4. At the same time, both of them could inhibit the protein expression of CYP3A4, as well as the effect degree of difference is bigger, which may be associated with its hydroxyl and the methoxyl isomerisation.